LOCUS SYNPBR322 4361 bp DNA circular SYN 30-SEP-2008
DEFINITION Cloning vector pBR322, complete sequence.
ACCESSION J01749 K00005 L08654 M10282 M10283 M10286 M10356 M10784 M10785
M10786 M33694 V01119
VERSION J01749.1
KEYWORDS ampicillin resistance; beta-lactamase; cloning vector; drug
resistance protein; origin of replication; plasmid; tetracycline
resistance.
SOURCE Cloning vector pBR322
ORGANISM Cloning vector pBR322
other sequences; artificial sequences; vectors.
REFERENCE 1 (bases 1 to 3; 3259 to 4361)
AUTHORS Sutcliffe,J.G.
TITLE Nucleotide sequence of the ampicillin resistance gene of
Escherichia coli plasmid pBR322
JOURNAL Proc. Natl. Acad. Sci. U.S.A. 75 (8), 3737-3741 (1978)
PUBMED 358200
REFERENCE 2 (bases 1 to 4361)
AUTHORS Sutcliffe,J.G.
TITLE Complete nucleotide sequence of the Escherichia coli plasmid pBR322
JOURNAL Cold Spring Harb. Symp. Quant. Biol. 43 (Pt 1), 77-90 (1979)
PUBMED 383387
REFERENCE 3 (bases 1500 to 2300)
AUTHORS Reed,R.R., Young,R.A., Steitz,J.A., Grindley,N.D. and Guyer,M.S.
TITLE Transposition of the Escherichia coli insertion element gamma
generates a five-base-pair repeat
JOURNAL Proc. Natl. Acad. Sci. U.S.A. 76 (10), 4882-4886 (1979)
PUBMED 388421
REFERENCE 4 (bases 2207 to 2265)
AUTHORS Covarrubias,L., Cervantes,L., Covarrubias,A., Soberon,X.,
Vichido,I., Blanco,A., Kupersztoch-Portnoy,Y.M. and Bolivar,F.
TITLE Construction and characterization of new cloning vehicles. V.
Mobilization and coding properties of pBR322 and several deletion
derivatives including pBR327 and pBR328
JOURNAL Gene 13 (1), 25-35 (1981)
PUBMED 6263753
REFERENCE 5 (bases 2000 to 2500)
AUTHORS Marians,K.J., Soeller,W. and Zipursky,S.L.
TITLE Maximal limits of the Escherichia coli replication factor Y
effector site sequences in pBR322 DNA
JOURNAL J. Biol. Chem. 257 (10), 5656-5662 (1982)
PUBMED 6279609
REFERENCE 6 (bases 1 to 80; 4151 to 4229; 4349 to 4361)
AUTHORS Brosius,J., Cate,R.L. and Perlmutter,A.P.
TITLE Precise location of two promoters for the beta-lactamase gene of
pBR322. S1 mapping of ribonucleic acid isolated from Escherichia
coli or synthesized in vitro
JOURNAL J. Biol. Chem. 257 (15), 9205-9210 (1982)
PUBMED 6178738
REFERENCE 7 (bases 4241 to 4343)
AUTHORS Van Dyke,M.W., Hertzberg,R.P. and Dervan,P.B.
TITLE Map of distamycin, netropsin, and actinomycin binding sites on
heterogeneous DNA: DNA cleavage-inhibition patterns with
methidiumpropyl-EDTA.Fe(II)
JOURNAL Proc. Natl. Acad. Sci. U.S.A. 79 (18), 5470-5474 (1982)
PUBMED 6291045
REFERENCE 8 (bases 584 to 709)
AUTHORS Peden,K.W. and Nathans,D.
TITLE Local mutagenesis within deletion loops of DNA heteroduplexes
JOURNAL Proc. Natl. Acad. Sci. U.S.A. 79 (23), 7214-7217 (1982)
PUBMED 6760191
REFERENCE 9 (bases 373 to 649)
AUTHORS Peden,K.W.
TITLE Revised sequence of the tetracycline-resistance gene of pBR322
JOURNAL Gene 22 (2-3), 277-280 (1983)
PUBMED 6307828
REFERENCE 10 (bases 132 to 181)
AUTHORS Watabe,H., Iino,T., Kaneko,T., Shibata,T. and Ando,T.
TITLE A new class of site-specific endodeoxyribonucleases. Endo.Sce I
isolated from a eukaryote, Saccharomyces cerevisiae
JOURNAL J. Biol. Chem. 258 (8), 4663-4665 (1983)
PUBMED 6300094
REFERENCE 11 (bases 368 to 581)
AUTHORS Livneh,Z.
TITLE Directed mutagenesis method for analysis of mutagen specificity:
application to ultraviolet-induced mutagenesis
JOURNAL Proc. Natl. Acad. Sci. U.S.A. 80 (1), 237-241 (1983)
PUBMED 6337373
REFERENCE 12 (bases 2627 to 2682; 2781 to 2828)
AUTHORS Mascharak,P.K., Sugiura,Y., Kuwahara,J., Suzuki,T. and Lippard,S.J.
TITLE Alteration and activation of sequence-specific cleavage of DNA by
bleomycin in the presence of the antitumor drug
cis-diamminedichloroplatinum(II)
JOURNAL Proc. Natl. Acad. Sci. U.S.A. 80 (22), 6795-6798 (1983)
PUBMED 6196777
REFERENCE 13 (bases 4276 to 4336)
AUTHORS Schultz,P.G. and Dervan,P.B.
TITLE Sequence-specific double-strand cleavage of DNA by
penta-N-methylpyrrolecarboxamide-EDTA X Fe(II)
JOURNAL Proc. Natl. Acad. Sci. U.S.A. 80 (22), 6834-6837 (1983)
PUBMED 6417654
REFERENCE 14 (bases 518 to 528)
AUTHORS Sutcliffe,J.G.
JOURNAL Unpublished
REFERENCE 15 (bases 2395 to 2495)
AUTHORS Fuller,R.S., Funnell,B.E. and Kornberg,A.
TITLE The dnaA protein complex with the E. coli chromosomal replication
origin (oriC) and other DNA sites
JOURNAL Cell 38 (3), 889-900 (1984)
PUBMED 6091903
REFERENCE 16 (bases 2729 to 2731)
AUTHORS Lathe,R., Kieny,M.P., Skory,S. and Lecocq,J.P.
TITLE Linker tailing: unphosphorylated linker oligonucleotides for
joining DNA termini
JOURNAL DNA 3 (2), 173-182 (1984)
PUBMED 6327214
REFERENCE 17 (bases 2729 to 2730)
AUTHORS Heusterspreute,M. and Davison,J.
TITLE Restriction site bank vectors. II. DNA sequence analysis of plasmid
pJRD158
JOURNAL DNA 3 (3), 259-268 (1984)
PUBMED 6086259
REFERENCE 18 (bases 2113 to 2186; 2348 to 2415)
AUTHORS Abarzua,P., Soeller,W. and Marians,K.J.
TITLE Mutational analysis of primosome assembly sites. I. Distinct
classes of mutants in the pBR322 Escherichia coli factor Y DNA
effector sequences
JOURNAL J. Biol. Chem. 259 (22), 14286-14292 (1984)
PUBMED 6209275
REFERENCE 19 (bases 2348 to 2415)
AUTHORS Soeller,W., Abarzua,P. and Marians,K.J.
TITLE Mutational analysis of primosome assembly sites. II. Role of
secondary structure in the formation of active sites
JOURNAL J. Biol. Chem. 259 (22), 14293-14300 (1984)
PUBMED 6150042
REFERENCE 20 (bases 1 to 4361)
AUTHORS Van Dyke,M.M. and Dervan,P.B.
TITLE Echinomycin binding sites on DNA
JOURNAL Science 225 (4667), 1122-1127 (1984)
PUBMED 6089341
REFERENCE 21 (sites)
AUTHORS Pouwels,P.H., Enger-Valk,B.E. and Brammar,W.J.
JOURNAL (in) CLONING VECTORS. Elsevier Scientific Publishing, Amsterdam
(1985)
REMARK Vector I-A-iv-1
REFERENCE 22 (bases 1 to 4361)
AUTHORS Watson,N.
TITLE A new revision of the sequence of plasmid pBR322
JOURNAL Gene 70 (2), 399-403 (1988)
PUBMED 3063608
REFERENCE 23 (sites)
AUTHORS Gilbert,W.
TITLE Obtained from VecBase 3.0
JOURNAL Unpublished
COMMENT On or before Apr 4, 2002 this sequence version replaced L08654.1,
gi:58257.
The circular sequence is numbered such that 0 is the middle of the
unique EcoRI site and the count increases first through the tet
genes, the pMB1 material, and finally through the Tn3 region.
Plasmid pBR322 contains ampicillin and tetracycline resistance
genes. The ampicillin resistance gene (amp-r) is a penicillin
beta-lactamase. Promoters P1 and P3 are for the beta-lactamase
gene. P3 is the natural promoter, and P1 is artificially created by
the ligation of two different DNA fragments to create pBR322. P2 is
in the same region as P1, but it is on the opposite strand and
initiates transcription in the direction of the tetracycline
resistance gene.
Mutational studies in the primosome assembly sites indicate four
types of mutations: Class I having no effect on the activities
elicited by the DNA site and the bases involved are probably
spacers; Class II requiring higher Mg-2+ concentrations than the
wild-type to be fully activated as factor Y ATPase effectors; Class
III co-inactivating both the ATPase effector and DNA replication
template activity of the site, indicating that they probably
represent essential contact points between factor Y and the DNA;
Class IV having a replication template activity intermediate that
of class III and class II mutant DNAs.
Specific sites within or near the origins of replication are
recognized by dnaA protein. Without dnaA binding to the origin of
replication chromosomal replication is not possible (15). pBR322
DNA contains two separate regions on opposite strands and close to
the origin of replication which, when in single-stranded form, can
act as effectors for the ATPase activity of E.coli replication
factor Y (5). Small fragments of DNA containing these sites when
cloned in an f1 phage vector act as origins of DNA replication
allowing the formation of complementary double-stranded DNA in
rifampicin-resistant, dna(B,G,C)-dependent fashion in vitro (5).
The biological activity of echinomycin is thought to be related to
the formation of complexes by intercalating with cellular DNA (20).
Complete source information:
Plasmid pBR322 from E.coli (2),(1),(3),(6),(11),(8),(5),(7),(12),
(13),(10),(9),(14),(18),(19),(15),(20),(16); pBR322 DNA in pXf3
(4).
The following data and their annotation were supplied by Will
Gilbert under the auspices of the Curator Program.
CROSSREFERENCE
#parent
GenBank(50):pSC101C, GenBank(50):Trn3
#offspring
VecBase(3):pBR325, VecBase(3):pBR327,
VecBase(3):pBR328,
VecBase(3):pAT153, VecBase(3):pUC7,
VecBase(3):pJRD158,
VecBase(3):PiVX, VecBase(3):PiAN7,
VecBase(3):pSP64,
VecBase(3):pSP65, VecBase(3):pGEM1,
VecBase(3):pGEM2,
VecBase(3):pGEM3, VecBase(3):pGEM4,
VecBase(3):pKK223,
VecBase(3):pLBU3, VecBase(3):pTrS3,
VecBase(3):pRSVNeo,
VecBase(3):pSV2Cat, VecBase(3):M13mp9,
VecBase(3):pHC79,
VecBase(3):pV34, VecBase(3):pKTH601,
VecBase(3):pKTH604,
VecBase(3):pKTH605, VecBase(3):pKTH606,
VecBase(3):YEp24,
VecBase(3):YIp5, VecBase(3):YRp17,
VecBase(3):pSP18,
VecBase(3):pSP19,
VecBase(3):pSP6T3, VecBase(3):pSP6T719,
VecBase(3):pT712,
VecBase(3):pT713, VecBase(3):pT7T318,
VecBase(3):pT7T319,
VecBase(3):pT7T3A18, VecBase(3):pT7T3A19,
VecBase(3):pEX1,
VecBase(3):pEX2, VecBase(3):pEX3,
VecBase(3):pCKSP6,
VecBase(3):pACYC177, VecBase(3):pKO1,
VecBase(3):pKO2,
VecBase(3):pKM1,
VecBase(3):pKM2, VecBase(3):pMBL1,
VecBase(3):pMBL604,
VecBase(3):pMC1511, VecBase(3):pMC1871,
VecBase(3):pAA37X,
VecBase(3):pUR278, VecBase(3):pUR288,
VecBase(3):pUR289,
VecBase(3):pUR290, VecBase(3):pUR291,
VecBase(3):pUR292,
VecBase(3):pUR222.
FEATURES Location/Qualifiers
source 1..4361
/organism="Cloning vector pBR322"
/mol_type="other DNA"
/db_xref="taxon:47470"
/tissue_lib="ATCC 31344, ATCC 37017"
/focus
source 1..1762
/organism="Plasmid pSC101"
/mol_type="other DNA"
/db_xref="taxon:2625"
misc_binding 24..27
/bound_moiety="echinomycin"
regulatory complement(27..33)
/regulatory_class="promoter"
/note="promoter P1 (6)"
misc_binding 39..42
/bound_moiety="echinomycin"
regulatory 43..49
/regulatory_class="promoter"
/note="promoter P2 (6)"
misc_binding 53..56
/bound_moiety="echinomycin"
misc_binding 67..70
/bound_moiety="echinomycin"
misc_binding 80..83
/bound_moiety="echinomycin"
gene 86..1276
/gene="tet"
CDS 86..1276
/gene="tet"
/codon_start=1
/transl_table=11
/product="tetracycline resistance protein"
/protein_id="AAB59735.1"
/translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASH
YGVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAG
RIVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLH
APFLAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTV
FFIMQLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGE
KQAIIAGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQG
QLQGSLAALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSR
ATST"
misc_feature complement(141..142)
/note="Endo.Sce I cleavage site coordinated with site at
base 146 (10)"
misc_feature 146..147
/gene="tet"
/note="Endo.Sce I cleavage site coordinated with site at
base 142 (10)"
misc_binding 411..414
/gene="tet"
/bound_moiety="echinomycin"
misc_difference 426
/gene="tet"
/note="conflict"
/citation=[11]
/replace=""
misc_binding 469..472
/gene="tet"
/bound_moiety="echinomycin"
old_sequence 526..528
/gene="tet"
/citation=[17]
repeat_region complement(1515..1519)
/note="gamma-delta insertion target sequence"
/rpt_type=direct
misc_feature 1636..1762
/note="from pSC101 (bp 1860-1986)"
repeat_region complement(1788..1792)
/note="gamma-delta insertion target sequence"
/rpt_type=direct
misc_difference 1891..1892
/note="conflict"
/citation=[23]
/replace="att"
old_sequence 1892..1893
/citation=[2]
/citation=[22]
regulatory 1905..1910
/regulatory_class="ribosome_binding_site"
regulatory 1905..1909
/regulatory_class="ribosome_binding_site"
/note="Shine-Dalgarno sequence"
misc_difference 1913..1914
/note="conflict"
/citation=[23]
/replace="caa"
old_sequence 1914..1915
/citation=[17]
CDS 1915..2106
/codon_start=1
/transl_table=11
/product="ROP protein"
/protein_id="AAB59736.1"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDH
ADELYRSCLARFGDDGENL"
misc_feature 2011..2167
/note="H-strand Y effector site"
/citation=[5]
repeat_region complement(2245..2249)
/note="gamma-delta insertion target sequence"
/rpt_type=direct
misc_feature complement(2351..2414)
/note="L-strand Y effector site"
/citation=[5]
misc_binding 2439..2447
/bound_moiety="dnaA"
rep_origin 2535
old_sequence 2729..2730
/note="revision according to (17)"
/citation=[2]
/citation=[17]
/replace="at"
old_sequence 2729
/citation=[17]
old_sequence 2730
/note="revision according to (16)"
/citation=[2]
/citation=[16]
/citation=[17]
/replace="t"
mobile_element 3148..4361
/mobile_element_type="transposon:Tn3"
repeat_region 3148..3185
/note="corresponds to one of the 38bp repeats found in Tn3
(bp 1-38 and complement (4920-4957))"
/rpt_type=inverted
gene complement(3293..4153)
/gene="bla"
CDS complement(3293..4153)
/gene="bla"
/note="E-286"
/codon_start=1
/transl_table=11
/product="beta-lactamase"
/protein_id="AAB59737.1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW"
sig_peptide complement(4085..4153)
/gene="bla"
mat_peptide complement(3296..4084)
/gene="bla"
/product="beta-lactamase"
regulatory complement(4161..4165)
/regulatory_class="ribosome_binding_site"
/note="Shine-Dalgarno sequence"
regulatory complement(4188..4194)
/regulatory_class="promoter"
/note="promoter P3 (6)"
misc_binding complement(4268..4271)
/bound_moiety="echinomycin"
misc_binding complement(4280..4283)
/bound_moiety="echinomycin"
misc_binding complement(4285..4288)
/bound_moiety="echinomycin"
misc_binding complement(4296..4299)
/bound_moiety="echinomycin"
misc_binding complement(4311..4314)
/bound_moiety="echinomycin"
misc_binding complement(4317..4320)
/bound_moiety="echinomycin"
misc_binding complement(4331..4334)
/bound_moiety="echinomycin"
BASE COUNT 983 a 1210 c 1134 g 1034 t
ORIGIN
1 ttctcatgtt tgacagctta tcatcgataa gctttaatgc ggtagtttat cacagttaaa
61 ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg
121 caccgtcacc ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt
181 gcgggatatc gtccattccg acagcatcgc cagtcactat ggcgtgctgc tagcgctata
241 tgcgttgatg caatttctat gcgcacccgt tctcggagca ctgtccgacc gctttggccg
301 ccgcccagtc ctgctcgctt cgctacttgg agccactatc gactacgcga tcatggcgac
361 cacacccgtc ctgtggatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac
421 aggtgcggtt gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca
481 cttcgggctc atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg
541 actgttgggc gccatctcct tgcatgcacc attccttgcg gcggcggtgc tcaacggcct
601 caacctacta ctgggctgct tcctaatgca ggagtcgcat aagggagagc gtcgaccgat
661 gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt
721 cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct
781 ctgggtcatt ttcggcgagg accgctttcg ctggagcgcg acgatgatcg gcctgtcgct
841 tgcggtattc ggaatcttgc acgccctcgc tcaagccttc gtcactggtc ccgccaccaa
901 acgtttcggc gagaagcagg ccattatcgc cggcatggcg gccgacgcgc tgggctacgt
961 cttgctggcg ttcgcgacgc gaggctggat ggccttcccc attatgattc ttctcgcttc
1021 cggcggcatc gggatgcccg cgttgcaggc catgctgtcc aggcaggtag atgacgacca
1081 tcagggacag cttcaaggat cgctcgcggc tcttaccagc ctaacttcga tcactggacc
1141 gctgatcgtc acggcgattt atgccgcctc ggcgagcaca tggaacgggt tggcatggat
1201 tgtaggcgcc gccctatacc ttgtctgcct ccccgcgttg cgtcgcggtg catggagccg
1261 ggccacctcg acctgaatgg aagccggcgg cacctcgcta acggattcac cactccaaga
1321 attggagcca atcaattctt gcggagaact gtgaatgcgc aaaccaaccc ttggcagaac
1381 atatccatcg cgtccgccat ctccagcagc cgcacgcggc gcatctcggg cagcgttggg
1441 tcctggccac gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct aggctggcgg
1501 ggttgcctta ctggttagca gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc
1561 tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg
1621 taaagtctgg aaacgcggaa gtcagcgccc tgcaccatta tgttccggat ctgcatcgca
1681 ggatgctgct ggctaccctg tggaacacct acatctgtat taacgaagcg ctggcattga
1741 ccctgagtga tttttctctg gtcccgccgc atccataccg ccagttgttt accctcacaa
1801 cgttccagta accgggcatg ttcatcatca gtaacccgta tcgtgagcat cctctctcgt
1861 ttcatcggta tcattacccc catgaacaga aatccccctt acacggaggc atcagtgacc
1921 aaacaggaaa aaaccgccct taacatggcc cgctttatca gaagccagac attaacgctt
1981 ctggagaaac tcaacgagct ggacgcggat gaacaggcag acatctgtga atcgcttcac
2041 gaccacgctg atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga cggtgaaaac
2101 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
2161 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc
2221 cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg
2281 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
2341 gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
2401 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
2461 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
2521 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
2581 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
2641 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
2701 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
2761 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
2821 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
2881 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
2941 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
3001 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
3061 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
3121 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
3181 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
3241 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
3301 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
3361 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
3421 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
3481 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
3541 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
3601 ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
3661 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
3721 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
3781 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
3841 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
3901 cggcgtcaac acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
3961 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
4021 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
4081 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
4141 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
4201 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
4261 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
4321 ataaaaatag gcgtatcacg aggccctttc gtcttcaaga a
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